Cloning, Expression and Characterization of a Lipase Gene from Marine Bacterium Pseudoalteromonas lipolytica SCSIO 04301  

作　　者：SU Hongfei MAI Zhimao ZHANG Si SU Hongfei;MAI Zhimao;ZHANG Si(Key Laboratory of Tropical Marine Bio-Resources and Ecology and Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P. R. China;University of Chinese Academy of Sciences, Beijing 100049, P. R. China)

机构地区：[1]Key Laboratory of Tropical Marine Bio-Resources and Ecology and Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, P. R. China [2]University of Chinese Academy of Sciences, Beijing 100049, P. R. China

出　　处：《Journal of Ocean University of China》2016年第6期1051-1058,共8页中国海洋大学学报（英文版）

基　　金：supported by the Administration of Ocean and Fisheries of Guangdong Province (GD2012D01-002);the ‘Strategic Priority Research Program’ of the Chinese Academy of Sciences (No.XDA10030400);the Natural Science Foundation of Guangdong Province, China (Grant Nos.2015A030310270 and 2016A 030313157)

摘　　要：Absract A lipase gene, 1ip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80kDa. Lip1233 was grouped into the lipase family X because it contained a highly conserved motif GHSLG. The recombinant enzyme was purified with Ni-NTA affinity chro- matography. The optimal temperature and pH value of Lip1233 were 45 ℃ and 8.0, respectively. It retained more than 70% of origi- nal activity after being incubated in pH ranging from 6.0 to 9.5 for 30min. It was stable when the temperature was below 45℃, but was unstable when the temperature was above 55℃. Most metal ions tested had no significant effect on the activity of Lip1233. Lip1233 remained more than original activity in some organic solvents at the concentration of 30% （v/v）. It retained more than 30% activity after incubated in pure organic solvents for 12 h, while in hexane the activity was nearly 100%. Additionally, Lip 1233 exhib- ited typical halotolerant characteristic as it was active under 4M NaC1. Lip1233 powder could catalyze efficiently the synthesis of fructose esters in hexane at 400C. These characteristics demonstrated that Lip1233 is applicable to elaborate food processing and organic synthesis.Absract A lipase gene, lip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80 k Da. Lip1233 was grouped into the lipase family X because it contained a highly conserved motif GHSLG. The recombinant enzyme was purified with Ni-NTA affinity chromatography. The optimal temperature and p H value of Lip1233 were 45℃ and 8.0, respectively. It retained more than 70% of original activity after being incubated in p H ranging from 6.0 to 9.5 for 30 min. It was stable when the temperature was below 45℃, but was unstable when the temperature was above 55℃. Most metal ions tested had no significant effect on the activity of Lip1233. Lip1233 remained more than original activity in some organic solvents at the concentration of 30%(v/v). It retained more than 30% activity after incubated in pure organic solvents for 12 h, while in hexane the activity was nearly 100%. Additionally, Lip1233 exhibited typical halotolerant characteristic as it was active under 4M Na Cl. Lip1233 powder could catalyze efficiently the synthesis of fructose esters in hexane at 40℃. These characteristics demonstrated that Lip1233 is applicable to elaborate food processing and organic synthesis.

关 键 词：LIPASE organic-solvent-tolerance HALOTOLERANCE fructose ester Pseudoalteromonas lipolytica SCSIO 04301 

分 类 号：Q78[生物学—分子生物学]